SUPERBROTH, broth, media

Thanks to those who replied.... I had 3 messages within the hour!!
I'll add them in here for anyone else who might be interested.
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Chris_Upton at darwin.biochem.ualberta.ca
Biochemistry Department
University of Alberta
Edmonton, AB T6H 2H7
Canada
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For plasmid preps (both large scale and mini) I habitually use
2xYT + 0.2% glucose. 2x YT is (per liter): 16 g Difco Tryptone
10 g Difco Yeast Extract
5 g NaCl
I confess I have never quantitated the difference in yield between this and
standard broth, but eyeball measurement has indicated it to be significant.
Typical yields for 10 ml minipreps are 50-100 ug for pUC-like plasmids. My best
1 liter prep was > 9 mg plasmid. We get different yields from different strains,
but in general we get better yields from "upscale" media. Whether it's worth
it is up to you and probably depends on what your plasmid copy number is.
Most of us would not use up 9 mg of plasmid in a century, so it may be overkill.
(However, we use pUC for restriction and transformation controls all the time -
my 9 mg only lasted about 2 years.)
Good luck,
Mike R.
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| Dr. Michael P. Ready | READY at UTBC01.CM.UTEXAS.EDU |
| Clayton Foundation Biochemical Institute | Phone: (512)471-3625 |
| University of Texas | FAX: (512)471-8696 |
| Austin, Texas 78737 | |
|__________________________________________|____________________________|
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Terrific Broth consists of the following components:
Solution A: 12g tryptone, 24g yeast extract, and 4 ml of glycerol
add water to 900 ml
Solution B: 0.17M KH2PO4, 0.72M K2HPO4
Before use, mix solution A and B at a ratio of 9:1.
Good Luck!
Song-Muh Jong
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Hi,
Bio101 sell a rich medium they call "Circlegrow" which appears to be
equivalent to TB (Terrific Broth). I use it for ssDNA preps from phagemids.
It seems to work OK.
Good Luck!
Chris Clegg